Benzothienyl aminoethyl ketones and their application in therapeutics

ABSTRACT

Compounds having the formula   &lt;IMAGE&gt;

The present invention has as its object new benzothienyl aminoethylketones, a method of preparation thereof and their application intherapeutics.

The compounds of the invention have the following general formula:##STR4## where: R₁ and R₂ each designate a saturated or unsaturatedaliphatic hydrocarbon having 1 to 4 carbon atoms or form together withthe nitrogen atom to which they are bound a heterocyclic radical chosenfrom the following: pyrrolidino, piperidino, perhydroazepino andmorpholino; and

Rx represents an acid compound chosen from the following:

Hydrochloric acid, bromhydric acid, sulphuric acid, phosphoric acid,boric acid;

Oxalic, maleic, malic, fumaric, citric, embonic, methane sulfonic,acetylsalicylic, nicotinic, parachlorophenoxyacetic orparachlorophenoxyisobutyric acid;

Methyl bromide, methyl iodide, ethyl bromide, butyl bromide, benzylbromide.

The method of the invention consists in condensing 2-acetylbenzothiophene of formula: ##STR5## with an amine of formula: ##STR6##according to Mannich's reaction.

The following preparation is given as an example to illustrate theinvention.

EXAMPLE 1-(benzothien-2'yl) 3-N-perhydroazepino propanone oxalate.Compound no. 1

A mixture of 8g of 2-acetyl benzothiophene, 6g of perhydroazepinehydrochloride and 3g of trioxymethylene in 30 ml of absolute ethanol, isheated to reflux for 4 hours. After dry vacuum evaporation, 100 ml of asaturated solution of sodium carbonate is added to the residue andextracted with diethyl ether. The ether phase is washed with a saturatedsolution of sodium chloride and dried on anhydrous sodium sulfate. Thesolvent is eliminated and the residue is dissolved in the minimum ofacetone. Three equivalents of oxalic acid in solution in acetone areadded and the obtained mixture is heated at reflux for 30 minutes. Thecrystals are centrifuged after cooling and recrystallised in a mixtureof methanol (1) and acetonitrile (1).

White crystals are thus obtained having a melting point of 200° C with ayield of 50%.

The compounds listed in the following table I were prepared according tothe same working method.

                                      TABLE I                                     __________________________________________________________________________     ##STR7##                                 (I)                                   compoundNo. of                                                                     ##STR8##    HX    (° C)pointMelting                                                           solventCrystallisation                                                                    Yield (%)                           __________________________________________________________________________           ##STR9##   Oxalic acid                                                                         118  Absolute ethanol (1) Ethyl ether                                                          60)                                  3                                                                                    ##STR10##  Citric acid                                                                         134  Acetonitrile                                                                              50                                   4                                                                                    ##STR11##  Oxalic acid                                                                         238  Methanol    50                                   5                                                                                    ##STR12##  Oxalic acid                                                                         186  Absolute ethanol                                                                          70                                   __________________________________________________________________________

The compounds of formula (I) were tested on laboratory animals andproved particularly active as blood-plaque aggregation inhibitors. Theyshowed also considerable activity as spasmolytic, analgesic,anti-inflammatory and coronary, cerebral and peripheral vasodilatatoryagents.

Furthermore, their toxicity was measured.

1. Acute toxicity

Acute toxicity of compounds of formula (I) was measured orally(esophageal probe) on mice deprived of food 18 hours before thebeginning of the test. The products were suspended in a solution ofdilute carboxymethyl cellulose. Calculation of the 50% lethal dose wascarried out according to the method of Miller and Tainter (Miller L. C.,Tainter M. L., proc. Soc. Exptl. Biol. Med. 1944, 57, 261-264).

The mortality rate was checked for seven days following the treatment.

Table II shows the 50% lethal doses (LD 50) in milligrammes perkilogramme of body weight.

                  TABLE II                                                        ______________________________________                                        acute toxicity                                                                No. of                                                                        compound    1        2      3      4     5                                    ______________________________________                                        LD 50       1200     960    1000   >1100 >1100                                (mg/kg/p.o.)                                                                  ______________________________________                                    

2. Activity on blood-plaque aggregation

The blood-plaques used in this study are human blood-plaques introducedinto the test as blood-plaque rich plasmas (B.P.R.P.). These latter areobtained as water-repellent material by slow centrifugation of humanblood samples.

The study of blood-plaque aggregation is made by means of anaggregatemeter with continuous agitation and graphic recording accordingto the conventional method (Prost R. S., Souverain C. H., Doumenc J.,Etude de l'aggregation plaquettaire a l'aide de l'agregametre de Mustard-- Coagulation, 1971, 4, 2, 145-151).

The aggregation inducing agents used are diphosphoric adenosine acid(D.P.A.), 1-(3',4'-di-hydroxyphenyl) 2-methylamino ethanol (known underthe name "adrenaline") and collagene. All these reactants are preparedfrom parent solutions diluted in Michaelis' buffer. Their concentrationin the test may vary according to the affinity of the blood-plaques.

An optimal concentration is thus sought by successive tests on the sameB.P.R.P.

The aggregation inhibiting activity was calculated by addition to theB.P.R.P. of the product to be tested before introduction of theaggregating agent.

The concentrations of the compounds of formula I (X) in the B.P.R.P. are2.5 × 10⁻⁵ g/ml for aggregation with D.P.A. and adrenaline and 2 × 10⁻⁵g/ml for collagene.

The reference product (R) is 2,6-bis (diethanolamino)4,8-dipiperidinopyrimido [5,4-d] pyrimidine (known under the trademark"Dipyridamole") used in the concentration of 5 × 10⁻⁴ g/ml foraggregation with D.P.A. and adrenaline and 2 × 10⁻⁴ g/ml for aggregationwith collagene.

The following table III shows the activity of some compounds of formulaI: a (X), the activity of the reference product: a (R) and the differentratio values: ##EQU1##

Furthermore, so as to have a better comparison between the activity ofthe compounds of the invention and that of the reference product, tableIII shows the ratio values: ##EQU2## (we have: LD 50(R) = 2150mg/kg/p.o.)

                                      TABLE III                                   __________________________________________________________________________    Blood-plaque aggregation inhibiting activity                                  No. of                                                                             D.P.A.      D.P.A. or Adrenaline                                                                      Adrenaline   Collagene                            com- pound tested                                                                  a(X) (%)                                                                         a(R) (%)                                                                         ##STR13##                                                                           ##STR14##   a(X) (%)                                                                         a(R) (%)                                                                         ##STR15##                                                                            a(X) (%)                                                                         a(R) (%)                                                                         ##STR16##                                                                           ##STR17##              __________________________________________________________________________    1    54 72 7.5 × 10.sup.-1                                                                  10.sup.-1                                                                               6 91  6 × 10.sup.-2                                                                 91 36 2.5     1.8 ×                                                                 10.sup.-1               2    74 53 1.4   1.1 × 10.sup.-1                                                                     98 96 1      74 42 1.75   2.25 ×                                                                 10.sup.-1               3    -- -- --    1.1 × 10.sup.-1                                                                     64 90 7 × 10.sup.-1                                                                  17 67 2.5 × 10.sup.-1                                                                2.15 ×                                                                 10.sup.-1               4    63 53 1.2   <1 × 10.sup.-1                                                                      89 80 1.1    91 42 2.2   <1.95 ×                                                                 10.sup.-1               5    -- -- --    <1 × 10.sup.-1                                                                      44 96 4.6 × 10.sup.-1                                                                45 36 1.25  <1.95 ×                                                                 10.sup.-1               __________________________________________________________________________

It appears that, taking into account the value of ratios: ##EQU3## thecompounds of the invention present an inhibiting activity onblood-plaque aggregation clearly superior to that of the referencesubstances.

3. Spasmolytic activity

Spasmolytic activity was studied on a rat's isolated duodenum maintainedin a survival medium of an aerated Tyrode solution heated to 38° C,according to the technique of Magnus (Magnus R. -- Archiv. ges.Physiol., 1905, 180, 1-71).

The contracting agent was barium chloride, the reference antagonistagent being papaverine hydrochloride (R). The average activity ofcompounds of formula I (X) compared to that of the reference agent wereexpressed by calculating the ratio of 50 efficient doses (ED 50)determined graphically on logarithmic paper: ##EQU4##

For example: a substance having a relative activity expressed by thefigure 2 has an activity equal to twice that of papaverinehydrochloride.

The results obtained shown in table IV show that the compounds of theinvention have an interesting spasmolytic activity.

                  TABLE IV                                                        ______________________________________                                        No. of   Spasmolytic activity                                                 compound tested                                                                        ED 50 (X) (g/ml)                                                                          ED 50 (R) (g/ml)                                                                           ##STR18##                                   ______________________________________                                        2         1.6 × 10.sup.-6                                                                    4.25 × 10.sup.-6                                                                    2.64                                         4        3.10 × 10.sup.-6                                                                    5.25 × 10.sup.-6                                                                    1.69                                         ______________________________________                                    

4. Analgesic and anti-inflammatory activity

The intraperitoneal injection of phenylparaquinone (P.P.Q.) to a mousecauses the appearance of a painful syndrome appearing within 5 minutesof the injection and disappearing after about 30 minutes (Siegmund E.A., Cadmus A., Lu G., J. Pharmacol. Exp. Therap. 1957, 119, 453).

This painful syndrome manifests itself in the animal by a series ofcharacteristic crises more or less close together and fleeting, duringwhich can be noted a twisting or stretching of the body, a hollowing ofthe sides and a stretching of the hind legs.

It is then possible to note the number of crises in a given interval oftime.

The preventive administration of analgesic or anti-inflammatorysubstances prevents the appearance of the crises and reduces thefrequence thereof.

The reference substance used in the tests (R) is the ester ofN-(7-chloro-4-quinolyl) anthrilic acid and 2,3-dihydroxy n. propanol(known under the trademark "Glafenine").

It is administered orally as a dose of 25 mg/kg.

The compounds of formula I (X) were administered digestively as a doseof 50 mg/kg.

The interval of time (t) chosen for calculating the analgesic activitywas between 10 and 15 minutes after injection of P.P.Q. which is theperiod of maximum frequency of the crises. This activity was expressedin the following manner;

n: the number of crises observed in controle mice;

n': the number of crises observed in mice previously treated with thereference substance (R);

n": the number of crises observed in mice having received substance (X)

The percentage activity of R is equal to: ##EQU5## The percentageactivity of X is equal to: ##EQU6## The activity of X in relation to Ris estimated by the ratio: ##EQU7## For example, a compound X for which:##EQU8## is calculated as 0.5 possesses an activity equal to half thatof the reference product.

The following table V shows the results observed.

                  TABLE V                                                         ______________________________________                                                 Analgesic and anti-inflammatory                                      No. of  activity (P.P.Q.)                                                     compound tested                                                                        a (X)    a (R)                                                                                  ##STR19##                                                                             ##STR20##                                  ______________________________________                                        2       36       71       0.51      5.2 × 10.sup.-2                     3       58       77       0.75      5 × 10.sup.-2                       4       25       75       0.33    <4.5 × 10.sup.-2                      5       54       71       0.76    <4.5 × 10.sup.-2                      ______________________________________                                    

It is clear that the reference product is more active than the compoundsof the invention. Nevertheless, these latter present an interestinganalgesic and anti-inflammatory activity since their activity representsa substantial fraction of that of the reference product which is anexcellent analgesic.

5. Vasodilatatory activity

The dilatatory activity as regards the smooth musculature of the vesselswas demonstrated on the isolated heart of a guinea-pig perfused with aLocke solution maintained at 37° C, after having provided the aorta witha cannula.

The coronary flow is recorded by means of an electronic devicecomprising a Fleish totaliser, before and after addition of substance Xto be studied (concentration: 1 × 10⁻⁵ g/ml), the reference productbeing 2,6-bis (diethanolamino) 4,8 dipiperidino pyrimido [5, 4-d]pyrimidine (known under the trademark "Dipyridamole") used in aconcentration of 1 × 10⁻⁵ g/ml.

The percentage of flow increase (P) is calculated at the climax of theactivity of X or R.

The following table shows the results observed.

                  TABLE VI                                                        ______________________________________                                        vasodilatatatory activity                                                     No. of compound                                                                            P (X)     P (R)                                                                                   ##STR21##                                    ______________________________________                                        1           66         76       0.87                                          2           64        162       0.4                                           ______________________________________                                    

These pharmacological results show the interest of compounds of formulaI in the treatment of coronary and cardiac inadequacies, pains ofinflammatory and other origins and spasms.

The new derivatives can be presented for oral, rectal or parenteraladministration in man and animals, particularly in association with theexcipients appropriate to these ways of administration.

Thus for example, they can be presented in the form of tablets, pills,gelules, suppositaries or injectable solutions.

The daily dose can, according to the case, be between 50 and 600 mg.

What we claim is:
 1. A compound having the formula (I): ##STR22## where:R₁ and R₂ each is alkyl or alkenyl having up to 4 carbon atoms or formtogether with the nitrogen atom to which they are bound pyrrolidino,piperidino or perhydroazepine; and RX is selected from the groupconsisting ofhydrochloric acid, bromhydric acid, sulphuric acid,phosphoric acid, boric acid, oxalic acid, maleic acid, malic acid,fumaric acid, citric acid, methane sulfonic acid, acetylsalicylic acid,nicotinic acid, parachlorophenoxyacetic acid,parachlorophenoxyisobutyric acid, methyl bromide, methyl iodide, ethylbromide, butyl bromide and benzyl bromide.
 2. A compound as claimed inclaim 1, 1-(benzothien-2'-yl)3-N-perhydroazepino propanone oxalate.
 3. Acompound as claimed in claim 1, 1-(benzothien-2'-yl)-3-dimethylaminopropanone oxalate.
 4. A compound as claimed in claim 1,1-(benzothien-2'-yl)-3-diallylamino propanone citrate.
 5. A compound asclaimed in claim 1, 1-(benzothien-2'-yl)-3-N-piperidino propanoneoxalate.
 6. A therapeutic composition useful as a blood plaqueaggregation inhibitor, a spasmolytic agent, an analgesic agent, ananti-inflammatory agent, a coronary vasodilatatory agent, a cerebralvasodilatatory agent or a peripheral vasodilatatory agent, comprising aneffective amount of a compound as claimed in claim 1 in combination witha pharmacologically acceptable carrier.